Study on Sustainable Recovery and Extraction of Polyhydroxyalkanoates (PHAs) Produced by Cupriavidus necator Using Waste Glycerol for Medical Applications

The current study shows that Cupriavidus necator has the ability to grow on waste glycerol as carbon source, and can synthesize a highly thermostable copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate). Batch cultivation on waste glycerol showed accumulation of 6.76 g L–1 biomass containing 4.84 g L–1 poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer with 3-hydroxyvalerate content of 24.6 mol%. A novel recovery strategy was developed for the extraction of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer from Cupriavidus necator using recyclable solvents, i.e., propylene carbonate, butyl acetate, isoamyl alcohol, and ethyl acetate. Propylene carbonate demonstrated the recovery efficiency of 90 % and polymer purity of 95 %, at 120 °C after 30 min. Ethyl acetate exhibited a higher efficiency than butyl acetate in terms of recovering the copolymer from cells. Ethyl acetate extraction demonstrated a recovery yield of 96 % and purity of 93 % at 100 °C. Efficacy of an anionic surfactant, linear alkylbenzene sulfonic acid (LAS), was also tested for extraction, and it showed maximum yield of 84 % and purity of 90 % at 80 °C and pH 5.0. Extraction of copolymer using these solvents could help in replacing generally used chlorinated toxic solvents, such as 1,2 dichloroethane and chloroform. Further, GPC, TGA and DSC analysis revealed that the thermo-physical properties were not significantly affected by the extraction method. However, the molecular weight distribution of the polymer showed a variation depending on the type of solvent used for extraction. Subsequently, endotoxins were removed efficiently to less than 5 EU g–1 of copolymer using alkali at optimized conditions of 6 h digestion time and 2.5 N NaOH concentration for medical applications

Atlantic salmon cardiac primary cultures:An in vitro model to study viral host pathogen interactions and pathogenesis

Development of Salmon Cardiac Primary Cultures (SCPCs) from Atlantic salmon pre-hatch embryos and their application as in vitro model for cardiotropic viral infection research are described. Producing SCPCs requires plating of trypsin dissociated embryos with subsequent targeted harvest from 24h up to 3 weeks, of relevant tissues after visual identification. SCPCs are then transferred individually to chambered wells for culture in isolation, with incubation at 15-22°. SCPCs production efficiency was not influenced by embryo's origin (0.75/ farmed or wild embryo), but mildly influenced by embryonic developmental stage (0.3 decline between 380 and 445 accumulated thermal units), and strongly influenced by time of harvest post-plating (0.6 decline if harvested after 72 hours). Beating rate was not significantly influenced by temperature (15-22°) or age (2-4 weeks), but was significantly lower on SCPCs originated from farmed embryos with a disease resistant genotype (F = 5.3, p<0.05). Two distinct morphologies suggestive of an ex vivo embryonic heart and a de novo formation were observed sub-grossly, histologically, ultra-structurally and with confocal microscopy. Both types contained cells consistent with cardiomyocytes, endothelium, and fibroblasts. Ageing of SCPCs in culture was observed with increased auto fluorescence in live imaging, and as myelin figures and cellular degeneration ultra-structurally. The SCPCs model was challenged with cardiotropic viruses and both the viral load and the mx gene expression were measurable along time by qPCR. In summary, SCPCs represent a step forward in salmon cardiac disease research as an in vitro model that partially incorporates the functional complexity of the fish heart

Enhancing the Production of Polyhydroxyalkanoate Biopolymer by Azohydromonas Australica Using a Simple Empty and Fill Bioreactor Cultivation Strategy

Polyhydroxyalkanoates (PHAs) are biodegradable polymers which are considered as an effective alternative for conventional plastics due to their mechanical properties similar to the latter. However, widespread use of these polymers is still hampered due to their high cost of production. This shortcoming could partly be resolved by obtaining high yields and productivity. In the present study, a drain-and-fill strategy of repeated-batch cultivation was adopted for the enhanced production of polyhydroxybutyrate PHB using Azohydromonas australica. In this strategy, 20 % (v/v) of the culture broth was removed from the reactor and supplemented with an equal volume of fresh medium. This strategy demonstrated a 3.3 fold and 1.8 fold increase in PHB concentration and productivity, respectively, as compared to batch cultivation. Repeated cultivation had also the benefit of avoiding non-productive time required for cleaning, refilling and sterilization of bioreactor during batch, thereby increasing the overall volumetric productivity and industrial importance of the process

Evaluation of role of piezo-surgical in mandibular impacted third molar removal: An original research

Aim: The purpose of the present research was to assess the role of piezo-surgical and its effectiveness against conventional techniques for removal of impacted mandibular third molar. Methodology: Study included 16 patients (8 male and 8 female) age 17 to 32 years treated in the period from 2019 to 2021. All patients had both mandibular molars impacted. One third molar was extracted using classical technique while the other one using a piezo device. Preoperative preparation was the same for all patients and included radiological analysis and verification of teeth by ortopan tomography (OPG) and CT. Patients were assessed one, seven and 14 days after the procedure. Results: After the extraction, all patients were followed for postoperative symptoms: pain, swelling and paraesthesia of the inferior alveolar nerve. The results confirmed advantages of piezosurgery in the removal of impacted mandibular third molars. Average duration of the intervention was 18 minutes with standard technique while the duration with piezo technique was 23 minutes. According to the visual-analogue scale (VAS) the average pain in the standard group was 9 whereas in the piezo group it was 6.&nbsp